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Background  Transgenic approaches provide a powerful tool for gene function investigations in plants. However, some legumes are still recalcitrant to current transformation technologies, limiting the extent to which functional genomic studies can be performed on.  Superroo t of  Lotus corniculatus  is a continuous root cloning system allowing direct somatic embryogenesis and mass regeneration of plants. Recently, a technique to obtain transgenic  L. corniculatus  plants from  Superroot -derived leaves through  A. tumefaciens- mediated transformation was described. However, transformation efficiency was low and it took about six months from gene transfer to PCR identification.    Results  In the present study, we developed an  A. rhizogenes -mediated transformation of  Superroot -derived  L. corniculatus  for gene function investigation, combining the efficient  A. rhizogenes -mediated transformation and the rapid regeneration system of  Superroot . The transformation system using  A. rhizogenes  K599 harbouring pGFPGUS Plus  was improved by validating some parameters which may influence the transformation frequency. Using stem sections with one node as explants, a 2-day pre-culture of explants, infection with K599 at OD 600  = 0.6, and co-cultivation on medium (pH 5.4) at 22°C for 2 days enhanced the transformation frequency significantly. As proof of concept,  Superroot -derived  L. corniculatus  was transformed with a gene from wheat encoding an Na + /H +  antiporter ( TaNHX2 ) using the described system. Transgenic  Superroot  plants were obtained and had increased salt tolerance, as expected from the expression of  TaNHX2 .    Conclusion  A rapid and efficient tool for gene function investigation in  L. corniculatus  was developed, combining the simplicity and high efficiency of the  Superroot  regeneration system and the availability of  A. rhizogenes -mediated transformation. This system was improved by validating some parameters influencing the transformation frequency, which could reach 92% based on GUS detection. The combination of the highly efficient transformation and the regeneration system of  Superroot  provides a valuable tool for functional genomics studies in  L. corniculatus .

Agrobacterium rhizogenes-mediated transformation of Superroot-derived Lotus corniculatus plants: a valuable tool for functional genomics